Summary of the invention
The invention provides a kind of CHLOROPHYLLINE salt compound and preparation method thereof, the present invention is that a kind of stable in properties, photodynamic activity are strong, little, the quality controllable novel photosensitive agent of skin phototoxicity, is used for cancer, infection and other needs the photodynamic therapy of the disease of optical radiation treatment.
The objective of the invention is to be achieved through the following technical solutions:
A kind of CHLOROPHYLLINE salt compound, general formula is as follows:
Wherein, R
1Be independently selected from C
1-18Alkyl, C
1-18Alkenyl, C
1-8Alkoxyl group and C
1-18Acyl group, it is randomly replaced by one or more substituting groups that are selected from halogen and hydroxyl; R
2Be Na
+, carboxylic acid sodium or glucoside; R
3Be H, carboxyl or carboxylic acid sodium.
In preferred embodiments, R
1Be independently selected from CH
2(CH
2)
4CH
3, R
2Be Na
+, carboxylic acid sodium or glucoside, R
3Be H or carboxyl.
The example of water-soluble CHLOROPHYLLINE salt compounds of the present invention includes, but are not limited to:
(I)
(II)
The objective of the invention is to be achieved through the following technical solutions:
The invention provides the method for preparing the CHLOROPHYLLINE salt compounds, may further comprise the steps:
I, pheophorbide is converted into acid porphyrin;
II, with described acid porphyrin derivatize, obtain the complexing acid porphyrin of deriving;
III, make the described complexing acid porphyrin of deriving obtain the CHLOROPHYLLINE salt compound with reaction in the medium that is selected from water and aqueous organic solution.
The miscible solvent of the alkali aqueous solution of the employing in step I, the II is:
One or more mixtures in lower alkane alcohols, polyalcohols, ring-type ethers, ketone, nitrile, amides, sulfoxide class, sulfone class, the gylcol ether.
Described lower alkane alcohols is one or more mixtures in methyl alcohol, ethanol, n-propyl alcohol, the Virahol; Described polyalcohols is ethylene glycol, 1, one or more mixtures in 2-propylene glycol, the glycerine;
Described ring-type ethers is one or more mixtures in tetrahydrofuran (THF), dioxane, the morpholine; Described ketone is acetone; Described nitrile is one or more mixtures in acetonitrile, the succinonitrile; Described amides is methane amide, N, one or more mixtures in the dinethylformamide; Described sulfone class is tetramethylene sulfone; Described sulfoxide class methyl-sulphoxide; Described gylcol ether is one or more mixtures in methyl glycol, glycol monoethyl ether, the glycol ether.
Reaction described in the step II can be carried out under the temperature of wide region, for example 0-90 ℃, preferred 10-65 ℃, more preferably 15-60 ℃, for example carries out under 15-25 ℃; According to used condition, the reaction times was generally dozens of minutes to several days in the step II, a few hours for example, for example 4-24 hour, for example 12 hours.
On the other hand, the invention provides compound as photosensitive drug.
On the other hand, the invention provides a kind of photosensitive drug for photodynamic therapy, it comprises compound of the present invention or mixture of the present invention, and randomly comprises one or more pharmaceutically useful vehicle.
On the other hand, the invention provides the purposes of CHLOROPHYLLINE salt compounds, namely be used for the treatment of as photosensitive drug or diagnosing malignant tumor, precancerous lesion or benign lesion.
On the other hand, the invention provides the method for the treatment of malignant tumour, precancerous lesion or benign lesion, it comprises compound of the present invention or mixture of the present invention or its pharmaceutical composition of using significant quantity to described individuality, and imposes the rayed of the specific wavelength of significant quantity.
The present invention has following beneficial effect compared to existing technology:
R of the present invention
3Substituting group be the form of carboxylate salt, water-soluble strong, be conducive to prepare follow-up pharmaceutical preparation; Absorbing wavelength of the present invention is greater than 650 nanometers; The present invention has the quantum yield of higher triplet state quantum yield and single low heavy attitude oxygen; The present invention has low dark toxicity, is conducive to HUMAN HEALTH; Selectivity height of the present invention; Structure of the present invention is clear and definite, stable performance; The present invention has good fat water compatibility.
Embodiment
Embodiment 1:
A kind of CHLOROPHYLLINE salt compound, general formula is as follows:
Wherein, R
1Be independently selected from C
1-18Alkyl, C
1-18Alkenyl, C
1-8Alkoxyl group and C
1-18Acyl group, it is randomly replaced by one or more substituting groups that are selected from halogen and hydroxyl; R
2Be Na
+, carboxylic acid sodium or glucoside; R
3Be H, carboxyl or carboxylic acid sodium.
In preferred embodiments, R
1Be independently selected from CH
2(CH
2)
4CH
3, R
2Be Na
+, carboxylic acid sodium or glucoside, R
3Be H or carboxyl.
[0023]The example of the water-soluble CHLOROPHYLLINE salt compounds of present embodiment includes but not limited to:
(I)
(II)
The preparation method of above-mentioned CHLOROPHYLLINE salt compounds may further comprise the steps:
I, pheophorbide is converted into acid porphyrin;
II, with described acid porphyrin derivatize, obtain the complexing acid porphyrin of deriving;
III, make the described complexing acid porphyrin of deriving obtain the CHLOROPHYLLINE salt compound with reaction in the medium that is selected from water and aqueous organic solution.
The miscible solvent of the alkali aqueous solution of the employing in step I, the II is:
One or more mixtures in lower alkane alcohols, polyalcohols, ring-type ethers, ketone, nitrile, amides, sulfoxide class, sulfone class, the gylcol ether.
Described lower alkane alcohols is one or more mixtures in methyl alcohol, ethanol, n-propyl alcohol, the Virahol; Described polyalcohols is ethylene glycol, 1, one or more mixtures in 2-propylene glycol, the glycerine;
Described ring-type ethers is one or more mixtures in tetrahydrofuran (THF), dioxane, the morpholine; Described ketone is acetone; Described nitrile is one or more mixtures in acetonitrile, the succinonitrile; Described amides is methane amide, N, one or more mixtures in the dinethylformamide; Described sulfone class is tetramethylene sulfone; Described sulfoxide class methyl-sulphoxide; Described gylcol ether is one or more mixtures in methyl glycol, glycol monoethyl ether, the glycol ether.
Reaction described in the step II can be carried out under the temperature of wide region, for example 0-90 ℃, preferred 10-65 ℃, more preferably 15-60 ℃, for example carries out under 15-25 ℃; According to used condition, in 0.5-120 hour reaction times, be preferably 4-24 hour, more preferably 12 hours in the step II.
On the other hand, present embodiment provides the compound as photosensitive drug.
On the other hand, present embodiment provides a kind of photosensitive drug for photodynamic therapy, and it comprises compound of the present invention or mixture of the present invention, and randomly comprises one or more pharmaceutically useful vehicle.
On the other hand, present embodiment provides the purposes of CHLOROPHYLLINE salt compounds, namely is used for the treatment of as photosensitive drug or diagnosing malignant tumor, precancerous lesion or benign lesion.
On the other hand, present embodiment provides the method for the treatment of malignant tumour, precancerous lesion or benign lesion, it comprises compound of the present invention or mixture of the present invention or its pharmaceutical composition of using significant quantity to described individuality, and imposes the rayed of the specific wavelength of significant quantity.
The R of the CHLOROPHYLLINE salt compound of present embodiment
3Substituting group be the form of carboxylate salt, water-soluble strong, be conducive to prepare follow-up pharmaceutical preparation; Absorbing wavelength is greater than 650 nanometers; Quantum yield with higher triplet state quantum yield and single low heavy attitude oxygen; Originally have low dark toxicity, be conducive to HUMAN HEALTH; Selectivity height of the present invention; Structure is clear and definite, stable performance; Has good fat water compatibility.
Among following examples 2-4, for those of ordinary skill in the art provides comprehensive and illustrative description, described how to prepare CHLOROPHYLLINE salt compounds of the present invention, and for the preparation of pharmaceutical composition, these embodiment limit the scope of the invention.Though in experiment, guarantee the accuracy of the data (for example quantity, temperature etc.) of use, also should consider some experimental error and deviation as far as possible.
Embodiment 2:
Present embodiment is raw material with silkworm excrement chlorophyll, and preparation 2-removes vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a sodium (1).
I, pheophorbide is converted into acid porphyrin:
A, preparation pheophorbide-a methyl esters:
200g silkworm excrement chlorophyll is dissolved in the methanol solution that 4000mL contains 5% sulfuric acid; 15-25 ℃ of lucifuge carried out stir process 12 hours under nitrogen protection again; Carry out filtration treatment again; Add water and methylene dichloride again and carry out extraction treatment; Again the organic phase water is carried out carrying out washing treatment; Carry out drying treatment, concentrating under reduced pressure processing again; Again the enriched material that obtains is carried out the silica gel column chromatography separating treatment, during described chromatographic separation is handled, eluent is sherwood oil and ethyl acetate, the volume ratio of described sherwood oil and ethyl acetate is 3:1, obtain blackish green solid A6.9g, wherein contain described pheophorbide-a methyl esters, productive rate is 85%.UV-vis?(CHCl
3)?λmax?:?407(1.0),?503?(0.03),?532?(0.02),?610?(0.02),?667?(0.39)?nm;?IR?(?KBr?)?ν:?2946-2843?(?C-H?),?1736?(?C=C?),?1640?(?C=N?),?1624?(?C=C?)?cm
-1;?
1H?NMR?(CDCl
3)?δ:?-1.68?(1H,?NH),?0.52(?1H,?NH),?1.70?(d,?J=7.5?Hz,?3H,?18-CH
3),?1.82?(t,?J=7.5Hz,?3H,?8
2-CH
3),?2.17-2.40?(m,?2H?17
1-CH
2),2.43-2.78?(m,2H,?17
2-H),?3.26,?3.40,?3.58,?3.67?(12H,?CH
3),?3.72?(q,?J=7.5?Hz,?2H,8
1-CH
2),?3.86?(s,?3H,?13
2-OCH
3),?4.20-4.36?(m,?1H,?17-H),?4.38-4.50?(m,?1H,?18-H),?6.24?(s,?1H,?13
2-H),6.16?(d,?J=11.5?Hz,?1H,?3
2b-H?),?6.29?(d,?J=17.5?Hz,?1H,?32a-H),?8.03?(m,?1H,?3
1-H),?8.57,?9.41,?9.53(?each?s,?1H,?meso-H?)。
II, with described acid porphyrin derivatize, obtain the complexing acid porphyrin of deriving;
A, preparation pyropheophorbide-a methyl esters:
The described blackish green solid A of 2.0g (3.3mmol) is dissolved with 200 mL glacial acetic acids, in stirring, carry out back flow reaction 4 h again; Carry out underpressure distillation again and handle, steam except most of acetic acid; Add 200 mL water again, (3 * 60 ml) carries out extraction treatment with methylene dichloride; Again organic phase is carried out drying back concentrating under reduced pressure; The gained enriched material being carried out silica gel column chromatography separates in the described chromatographic separation processing again, eluent is sherwood oil and ethyl acetate, the volume ratio of described sherwood oil and ethyl acetate is 3:1, obtain blackish green solid B (1.5g, 2.80mmol), wherein contain described pyropheophorbide-a methyl esters, productive rate is 85%.UV-vis?(CHCl
3)?λmax:?411?(1.0),?669?(0.39)?nm;IR(?KBr?)?ν:?2935-2846(?C-H?),?1736?(?C=C?),?1643?(?C=N?),?1619?(?C=C?)?cm
-1;?
1H?NMR?(CDCl
3)?δ:?-1.68?(1H,?NH),?0.49?(1H,?NH),1.69?(d,?J=7.5?Hz,?3H,?18-CH
3),?1.80?(t,?J=7.5?Hz,?3H,?8
2-CH
3),?2.14-2.38?(m,?2H?17
1-CH
2),?3.24,3.42,?3.66,?3.62(12H,?CH3),2.41-2.82?(m,2H,?17
2-CH
2),?3.70?(q,?J=7.5?Hz,?2H,?8
1-CH2),?4.17-4.38?(m,?1H,?17-H)?4.40-4.53?(m,?1H,?18-H),?5.15?(d,?J=20.0?Hz,?1H,?13
2-H),?5.26?(1H,?13
2-H),?6.19-?6.26?(m,?2H,?3
2-H),?8.00?(m,?1H,?3
1-H),?8.58,?9.45,?9.56?(?s,?1H,?meso-H?)。
B, preparation 2-remove vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a methyl esters:
With the described blackish green solid B of 500mg (0.93mmol) with the dissolving of 30% Hydrogen bromide glacial acetic acid (12.5 milliliters), then at 15-25 ℃ of stirring reaction 3 h; Carry out vacuum decompression again and handle, desolventizing gets and mainly contains the solid that 2-removes vinyl-2-(1-bromotrifluoromethane)-pyropheophorbide-a methyl esters; Above-mentioned mainly containing added 7.5 milliliters of n-hexyl alcohols in the solid that 2-removes vinyl-2-(1-bromotrifluoromethane)-pyropheophorbide-a methyl esters, add 60 milliliters of anhydrous methylene chlorides and 200 milligrams of Anhydrous potassium carbonates again; Reaction added 500 milliliters of methylene dichloride after 1 hour under nitrogen protection again; Again with the organic layer evaporate to dryness; Carry out the alumina column chromatography separating treatment again, during described chromatographic separation is handled, eluent is sherwood oil and acetone, the volume ratio of described sherwood oil and acetone is 4:1, namely get target product, wherein contain described 2-and go that vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a methyl esters productive rate is 55%.UV-vis?(CHCl
3)λmax:?410?(1.00),?505?(0.10),?536?(0.10),?667?(0.45)?nm;?IR?(KBr)?v:?2985-2870?(C-H),?1742,1708?(C=O),?1640?(C=N),?1620?(C=C)?cm
-1;
1H?NMR?(CDCl
3)?δ:?9.70,?9.53,?8.51?(each?s,?each?1H,?meso-H),?6.25-6.43?(m,?1H,3
1-H),?6.25?(s,1H,?13
2-H),?4.45-4.49?(?1H,?17-H),?4.15-4.26?(1H,?18-H),?3.88,?3.69,?3.55,?3.40,?3.23(each?s,?each?3H,?CH
3),?3.70?(q,?J=7.5?Hz,?8
1-CH
2),?2.18?(d,?J?=?7.0?Hz,?3
1-CH3),?2.00-2.70?(4H,?17-CH
2),?1.81?(t,?J=7.0?Hz,?8-CH
3),?1.73?(d,?J=7.5?Hz,?18-CH
3),?0.50,?-1.70?(each?br?s,?each1H,?NH)。
III, make the described complexing acid porphyrin of deriving obtain the CHLOROPHYLLINE salt compound with reaction in the medium that is selected from water and aqueous organic solution:
A, preparation 4,2-remove vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a sodium:
Make its whole dissolvings with adding 20 ml tetrahydrofuran (THF)s in the described target product of 100mg, add 4 ml, 0.2 mol/L aqueous sodium hydroxide solution again, place again and spend the night; Carry out reduced pressure treatment again, desolventizing obtains reaction product; Described reaction product is packed in the apparatus,Soxhlet's, and acetone refluxes, and gained acetone phegma evaporate to dryness namely gets target compound (1):
Each raw material, reagent and equipment are the market conventional products in the present embodiment.
Embodiment 3:
Present embodiment is raw material with silkworm excrement chlorophyll, and preparation 2-removes vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a disodium (2).
I, pheophorbide is converted into acid porphyrin:
A, preparation pheophorbide-a methyl esters:
100g silkworm excrement chlorophyll is dissolved in the methanol solution that 2000mL contains 5% sulfuric acid; Carried out stir process 12 hours in nitrogen protection 15-25 ℃ lucifuge again; Carry out filtration treatment again; Add water and methylene dichloride again and carry out extraction treatment; Again the organic phase water is carried out carrying out washing treatment; Carry out drying treatment, concentrating under reduced pressure processing again; Again the enriched material that obtains is carried out the silica gel column chromatography separating treatment, during described chromatographic separation is handled, eluent is sherwood oil and ethyl acetate, the volume ratio of described sherwood oil and ethyl acetate is 3:1, obtain blackish green solid A3.50g, wherein contain described pheophorbide-a methyl esters, productive rate is 85%.UV-vis?(CHCl
3)?λmax?:?407(1.0),?503?(0.03),?532?(0.02),?610?(0.02),?667?(0.39)?nm;?IR?(?KBr?)?ν:?2946-2843?(?C-H?),?1736?(?C=C?),?1640?(?C=N?),?1624?(?C=C?)?cm
-1;?
1H?NMR?(CDCl
3)?δ:?-1.68?(1H,?NH),?0.52(?1H,?NH),?1.70?(d,?J=7.5?Hz,?3H,?18-CH
3),?1.82?(t,?J=7.5Hz,?3H,?8
2-CH
3),?2.17-2.40?(m,?2H?17
1-CH
2),2.43-2.78?(m,2H,?17
2-H),?3.26,?3.40,?3.58,?3.67?(12H,?CH
3),?3.72?(q,?J=7.5?Hz,?2H,8
1-CH
2),?3.86?(s,?3H,?13
2-OCH
3),?4.20-4.36?(m,?1H,?17-H),?4.38-4.50?(m,?1H,?18-H),?6.24?(s,?1H,?13
2-H),6.16?(d,?J=11.5?Hz,?1H,?3
2b-H?),?6.29?(d,?J=17.5?Hz,?1H,?32a-H),?8.03?(m,?1H,?3
1-H),?8.57,?9.41,?9.53(?each?s,?1H,?meso-H?)。
II, with described acid porphyrin derivatize, obtain the complexing acid porphyrin of deriving:
A, 2,2-go the preparation of vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a dimethyl ester:
With the Hydrogen bromide glacial acetic acid dissolving of the described blackish green solid A of 1.0 g (0.93mmol) with 25ml30%, then at stirring reaction 3 h; Carry out vacuum decompression again and handle, desolventizing gets and mainly contains the solid that 2-removes vinyl-2-(1-bromotrifluoromethane)-pheophorbide-a dimethyl ester; Add 15 milliliters of n-hexyl alcohols in the solid that 2-removes vinyl-2-(1-bromotrifluoromethane)-pheophorbide-a dimethyl ester to above-mentioned mainly containing again, add 100 milliliters of anhydrous methylene chlorides and 500 milligrams of Anhydrous potassium carbonates again; Reaction added 1000 milliliters of methylene dichloride, with the organic layer evaporate to dryness after 1 hour under nitrogen protection again; Carry out the alumina column chromatography separating treatment again, during described chromatographic separation is handled, eluent is sherwood oil and acetone, the volume ratio of described sherwood oil and acetone is 4:1, namely get target product, wherein contain described 2,2-and remove vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a dimethyl ester, productive rate is 60%.
UV-vis?(CHCl
3)λmax:?410?(1.00),?505?(0.10),?536?(0.10),?667?(0.45)?nm;?IR?(KBr)?v:?2985-2870?(C-H),?1742,1708?(C=O),?1640?(C=N),?1620?(C=C)?cm
-1;
1H?NMR?(CDCl
3)?δ:?9.70,?9.53,?8.51?(each?s,?each?1H,?meso-H),?6.25-6.43?(m,?1H,3
1-H),?6.25?(s,1H,?13
2-H),?4.45-4.49?(?1H,?17-H),?4.15-4.26?(1H,?18-H),?3.88(s,?6?H,?CH
3),?3.69,?3.55,?3.40,?3.23(each?s,?each?3H,?CH
3),?3.70?(q,?J=7.5?Hz,?8
1-CH
2),?2.18?(d,?J?=?7.0?Hz,?3
1-CH3),?2.00-2.70?(4H,?17-CH
2),?1.81?(t,?J=7.0?Hz,?8-CH
3),?1.73?(d,?J=7.5?Hz,?18-CH
3),?0.50,?-1.70?(each?br?s,?each1H,?NH)。
III, make the described complexing acid porphyrin of deriving obtain the CHLOROPHYLLINE salt compound with reaction in the medium that is selected from water and aqueous organic solution:
A, 3,2-go the preparation of vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a disodium (2):
Make its whole dissolvings with adding 40 ml tetrahydrofuran (THF)s in the described target product of 200mg, add 10 ml, 0.2 mol/L aqueous sodium hydroxide solution again, place again and spend the night; Carry out reduced pressure treatment again, desolventizing obtains reaction product; Described reaction product is packed in the apparatus,Soxhlet's, and acetone refluxes, and gained acetone phegma evaporate to dryness namely gets target compound (II):
Each raw material, reagent and equipment are the market conventional products in the present embodiment.
Embodiment 4:
Present embodiment is that the detected result of embodiment 2, embodiment 3: 2-goes vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a sodium (1) and 2-to go vinyl-2-(the own hydroxyethyl of the 1-)-pheophorbide-photosensitization of a disodium (2) in the acellular system.
The photosensitization of compound judges that this compound is as the possibility of photo-dynamical medicine in heavy water, reference [Xu Deyu etc., the The 2nd Army Medical College journal, 1988,9(5): 435 and Li Yixin etc., Acta Biophysica Sinica, 1985,1(4): 257] described method, having compared 2-goes vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a sodium (1) and 2-to go vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a disodium (2) to the sensitization effect of NADPH photooxidation, to the results are shown in following table in heavy water.
According to The above results, 2-goes vinyl-2-(the own hydroxyethyl of 1-)-pyropheophorbide-a sodium (1) and 2-to go the photooxidation of vinyl-2-(the own hydroxyethyl of 1-)-pheophorbide-a disodium (2) all to be better than HpD.